- -

Evaluation of PCR and non-radioactive molecular hybridization techniques for the routine diagnosis of Tomato leaf curl new delhi virus, Tomato yellow leaf curl virus and tomato yellow leaf curl Sardinia virus

RiuNet: Institutional repository of the Polithecnic University of Valencia

Share/Send to

Cited by

Statistics

Evaluation of PCR and non-radioactive molecular hybridization techniques for the routine diagnosis of Tomato leaf curl new delhi virus, Tomato yellow leaf curl virus and tomato yellow leaf curl Sardinia virus

Show simple item record

Files in this item

dc.contributor.author Alfaro Fernández, Ana Olvido es_ES
dc.contributor.author Sanchez Navarro, Jesus Angel es_ES
dc.contributor.author Landeira, Micaela es_ES
dc.contributor.author Font San Ambrosio, Maria Isabel es_ES
dc.contributor.author Hernández Llopis, Desamparados es_ES
dc.contributor.author Pallás Benet, Vicente es_ES
dc.date.accessioned 2017-12-21T13:06:34Z
dc.date.available 2017-12-21T13:06:34Z
dc.date.issued 2016 es_ES
dc.identifier.issn 1125-4653 es_ES
dc.identifier.uri http://hdl.handle.net/10251/93285
dc.description.abstract [EN] The begomovirus Tomato leaf curl New Delhi virus (ToLCNDV) has been reported as a causal agent of leaf curl disease in tomato and other solanaceous crops and, more recently, affecting different cucurbitaceous crops. ToLCNDV was first detected in Asia and recently in Europe, in 2013. In the present analysis, we have evaluated the PCR and the non-radioactive nucleic acids spot hybridization (NASH) techniques together with two nucleic acids extraction protocols, for the routine diagnosis of ToLCNDV and its discrimination from the closely related Tomato yellow leaf curl Sardinia virus (TYLCSV) and Tomato yellow leaf curl virus (TYLCV). A protocol, designed to extract only the DNA, gave the best results with the PCR technique meanwhile the use of silica, which favors total nucleic acids extraction, was the best extraction protocol for the NASH. All allowed the specific detection of ToLCNDV but only some of the general begomovirus primers allowed the detection of all three viruses. The two ToLCNDV riboprobes analyzed by NASH, targeting the replicase and the coat protein genes, respectively, detected the virus with no cross-reaction with the TYLCV and TYLCSV infected extracts, obtaining a better detection limit with the replicase riboprobe. Direct comparison between the PCR and NASH techniques by the analysis of 42 field samples, revealed a good correlation between the two techniques. In addition, some samples were detected only by NASH due the presence of PCR inhibitors. The use of the PCR and NASH for the routine diagnosis of ToLCNDV is discussed. es_ES
dc.description.sponsorship We thank L. Corachan for her excellent technical assistance. This work was supported by grants BIO2014-54862-R from the Spanish granting agency DGICYT and SP20140770 funded by Spanish Ministry of Education, Culture and Sports (MECD) as a part of the Program of the Campus of International Excellence, which belongs to the Program of Assessing and Combined Resources R&D of VLC/CAMPUS2014. Authors want to thank Dr. P. Martinez-Culebras for the design of PGI/PGII primers included in Font (2003).
dc.language Inglés es_ES
dc.publisher EDIZIONE ETS, PISA, ITALY es_ES
dc.relation MINECO/BIO2014-54862-R es_ES
dc.relation UPV/SP20140770 es_ES
dc.relation.ispartof JOURNAL OF PLANT PATHOLOGY es_ES
dc.rights Reserva de todos los derechos es_ES
dc.subject Molecular detection methods es_ES
dc.subject Dig-RNA probe es_ES
dc.subject Begomovirus es_ES
dc.subject Routine diagnosis es_ES
dc.subject.classification PRODUCCION VEGETAL es_ES
dc.title Evaluation of PCR and non-radioactive molecular hybridization techniques for the routine diagnosis of Tomato leaf curl new delhi virus, Tomato yellow leaf curl virus and tomato yellow leaf curl Sardinia virus es_ES
dc.type Artículo es_ES
dc.identifier.doi 10.4454/JPP.V98I2.003 es_ES
dc.rights.accessRights Cerrado es_ES
dc.contributor.affiliation Universitat Politècnica de València. Instituto Universitario Mixto de Biología Molecular y Celular de Plantas - Institut Universitari Mixt de Biologia Molecular i Cel·lular de Plantes es_ES
dc.contributor.affiliation Universitat Politècnica de València. Instituto Agroforestal Mediterráneo - Institut Agroforestal Mediterrani es_ES
dc.contributor.affiliation Universitat Politècnica de València. Departamento de Ecosistemas Agroforestales - Departament d'Ecosistemes Agroforestals es_ES
dc.description.bibliographicCitation Alfaro Fernández, AO.; Sanchez Navarro, JA.; Landeira, M.; Font San Ambrosio, MI.; Hernández Llopis, D.; Pallás Benet, V. (2016). Evaluation of PCR and non-radioactive molecular hybridization techniques for the routine diagnosis of Tomato leaf curl new delhi virus, Tomato yellow leaf curl virus and tomato yellow leaf curl Sardinia virus. JOURNAL OF PLANT PATHOLOGY. 98(2):245-254. doi:10.4454/JPP.V98I2.003 es_ES
dc.description.accrualMethod S es_ES
dc.relation.publisherversion http://dx.doi.org/10.4454/JPP.V98I2.003 es_ES
dc.description.upvformatpinicio 245 es_ES
dc.description.upvformatpfin 254 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion es_ES
dc.description.volume 98 es_ES
dc.description.issue 2 es_ES
dc.relation.pasarela S\323470 es_ES
dc.contributor.funder Universitat Politècnica de València es_ES
dc.contributor.funder Ministerio de Economía, Industria y Competitividad es_ES


This item appears in the following Collection(s)

Show simple item record